Scaffold for bone and tissue repair in mammals

ABSTRACT

A mammalian tissue scaffold and method for making a tissue scaffold including a rigid scaffold body of biocompatible glass fibers bonded together and in special alignment to define open channels within the scaffold to allow fluid flow into and within the scaffold.

This is a continuation-in-part application based on U.S. applicationSer. No. 12/354,313 filed Jan. 15, 2009, issued Jan. 15, 2013 as U.S.Pat. No. 8,353,966, the entire disclosure of which is incorporated byreference.

FIELD OF THE INVENTION

This invention relates to a biocompatible scaffold for implantation intomammals to facilitate bone and tissue repair, regeneration, andproliferation.

BACKGROUND OF THE INVENTION

The use of tissue scaffold to facilitate the repair and regrowth of boneand other tissue is known in the art, for example, from U.S. Pat. No.7,338,517 which discloses an implantable scaffold with biopolymerfibrils aligned in helical patterns of opposite directions.

U.S. Pat. No. 7,416,564 describes a bone scaffold made from porousceramic substrate of a material such as zirconia which is then coatedwith a fluorapatite layer and a hydroxyapatite layer. The poroussubstrate was prepared by dipping a foam template into a slurry. Theporosity is aligned irregularly and randomly and does not runcontinuously along a longitudinal axis from one end of the object to theother end. The fabrication requires repeated dipping and drying.

SUMMARY OF THE INVENTION

Briefly, therefore, the invention is directed to a tissue scaffoldcomprising a rigid scaffold body having a scaffold central axis, ascaffold transverse dimension, and a scaffold lengthwise dimension whichis greater than the scaffold transverse dimension, the scaffold bodyhaving a compressive strength between about 20 and about 250 MPa andcomprising biocompatible inorganic glass fibers each having a fibertransverse dimension and a fiber lengthwise dimension which is at leastabout 10 times the fiber transverse dimension; wherein each of thefibers has a diameter between about 20 and about 5000 microns; whereinthe fibers are bonded together; and wherein at least about 75 vol % ofthe fibers extend generally in the direction of the scaffold centralaxis, are generally free of helical orientation about the scaffoldcentral axis, and are arranged to define open channels within thescaffold which allow fluid flow into and lengthwise within the scaffold.

In another aspect the invention is directed to a tissue scaffoldcomprising a rigid scaffold body having a central axis, a scaffoldtransverse dimension, and a scaffold lengthwise dimension which isgreater than the scaffold transverse dimension, the scaffold body havinga compressive strength between about 20 and about 250 MPa andcomprising: biocompatible inorganic glass fibers each having a fibertransverse dimension and a fiber lengthwise dimension which is at leastabout 10 times the fiber transverse dimension; wherein the fibers arebonded together; wherein each of the fibers has a diameter between about20 and about 5000 microns; and wherein at least about 75 vol % of thefibers extend generally parallel to the scaffold central axis, and arearranged to define open channels within the scaffold which allow fluidflow into and lengthwise within the scaffold.

The invention is also directed to a tissue scaffold comprising ascaffold body having a central axis, a scaffold transverse dimension,and a scaffold lengthwise dimension which is greater than the scaffoldtransverse dimension, the scaffold body comprising: biocompatibleinorganic glass fibers each having a fiber transverse dimension and afiber lengthwise dimension which is at least about 10 times the fibertransverse dimension; wherein each of the fibers has a diameter betweenabout 20 and about 5000 microns; wherein the fibers are bonded together;wherein at least about 75 vol % of the fibers extend generally in thedirection of the scaffold central axis, and are arranged to define openchannels lengthwise through a core within the scaffold, which channelsallow fluid flow into and lengthwise within the scaffold.

In another aspect the invention is directed to a method for making atissue scaffold comprising: heating inorganic biocompatible glass fibersin a mold to a temperature where the fibers partially fuse to each otherto form a rigid scaffold body having a scaffold central axis, a scaffoldtransverse dimension, and a scaffold lengthwise dimension which isgreater than the scaffold transverse dimension, the scaffold bodyhaving, a compressive strength between about 20 and about 250 MPa;wherein the biocompatible glass fibers each has a fiber transversedimension and a fiber lengthwise dimension which is at least about 10times the fiber transverse dimension; wherein the fibers have a diameterbetween about 20 and about 5000 microns; and wherein at least about 75vol % of the fibers extend generally in the direction of the scaffoldcentral axis, are generally free of helical orientation about thescaffold central axis, and are arranged to define open channels withinthe scaffold which allow fluid flow into and lengthwise within thescaffold.

Other objects and features of the invention are in part apparent and inpart pointed out hereinafter.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A is an optical micrograph of a transverse cross section of ascaffold of the invention formed by heating fibers at 700° C. for arelatively shorter time than with the scaffold in FIGS. 1B and 1C.

FIGS. 1B and 1C are SEM photographs of a transverse cross section of thescaffold of the invention formed by heating fibers at 700° C. for arelatively longer time than with the scaffold in FIG. 1A.

FIG. 2 is a photograph of a lengthwise cross section of the scaffold ofthe invention showing lengthwise and parallel alignment of fibers.

FIGS. 3A, 3B, and 3C are a series of photographs showing themanufacturing progression of a unidirectional scaffold from loose fibersto a self-bonded scaffold of the invention.

FIGS. 4A and 4B are photographs of a segment of scaffold of theinvention after immersion in an osteoblast cell culture and MTTlabeling.

FIGS. 5 and 6 are graphical plots of open porosity and compressivestrength data of scaffolds of the invention.

FIGS. 7A and 7B are photographs of a scaffold of the invention next to achicken bone.

FIGS. 8A, 8B, 8C, 8D, 9A, 9B, 9C, and 9D are still frames extracted fromvideos taken of experiments described in the working examples.

FIGS. 10, 12, and 13 are schematic depictions of alternative reinforcedscaffold embodiments of the invention.

FIG. 11 is a photograph of a scaffold according to the schematic of FIG.10.

FIGS. 14, 16, 18, 19 and 21 are a schematic end view depictions ofalternative scaffold embodiments.

FIGS. 15, 17, and 20 are photographs of alternative scaffoldembodiments.

DESCRIPTION OF PREFERRED EMBODIMENTS

The tissue scaffold of the present invention is prepared from fiberswhich are aligned so that a majority of the fibers are substantiallyaligned in a parallel direction. The scaffold is prepared by placing andorienting fibers in a unidirectional manner in a mold. The fibers in themold are heated to a temperature where the fibers soften and bondtogether. In one preferred embodiment, the fibers are self bonded in thesense that no adhesive, braze, or other external bonding agent is usedfor bonding. An alternative embodiment employs a biocompatible agent oradhesive to facilitate bonding, such that the fibers are not selfbonded, at least in part. Upon cooling, the assemblage of bonded fibersis sufficiently rigid and strong that the assemblage can be removed fromthe mold and handled. The scaffold is sufficiently rigid that it can beimplanted into a mammal where it facilitates the repair and regenerationof hard tissue such as bone (including cortical and cancellous) or softtissue such as muscle, cartilage, skin, organ, or other hard or softtissue.

The orientation of the fibers in a lengthwise direction in the selfbonded scaffold provides lengthwise channels (or connected pores) amongthe fibers, which channels provide for uptake into the scaffold of stemcells, growth factors, medicines, red blood cells and other bodilyfluids and components carried in bodily fluids. The fibers are arrangedto define channels within the scaffold which facilitate fluid flow intoand lengthwise within the scaffold from one end to the other end. Theorientation also provides for channels in a transverse directiongenerally perpendicular to the lengthwise direction of the orientedfibers, to facilitate uptake of fluids from the outer surface of theinterior or core of the scaffold. These longitudinal and transversechannels exert significant capillary forces on a liquid which cause theliquid to be drawn into the scaffold. This capillary action facilitatesthe distribution of these fluids and components relatively uniformlythrough the scaffold and enables fluids to flow from one end of thescaffold to the other or to enter the scaffold from its surface andtransmit the liquid to its ends.

The invention in one embodiment employs fibers having a diameter, priorto molding and softening, between about 20 and about 5000 microns, suchas between about 50 and about 5000 microns. In one embodiment thescaffold is prepared from fibers having diameters between about 100 andabout 450 microns, such as between about 100 and about 300 microns. Inan alternative embodiment, the scaffold is prepared from fibers havingdiameters up to about 3000 or 5000 microns (3 to 5 mm), which can bedeemed more akin to rods than fibers in some contexts, but for purposesof the discussion of this invention fall within the definition of“fibers.” FIG. 1A is an optical micrograph of a cross section of ascaffold of the invention showing the self-bonded fibers and pores afterheating the fibers at 700° C. for 15 minutes. FIGS. 1B and 1C, which areSEM photographs of a transverse cross section of a different scaffold ofthe invention having undergone a greater degree of softening and bondingthan the scaffold of FIG. 1A, show that after molding and joining to agreater degree (heating at 700° C. for 45 minutes), the transverse crosssection of each fiber is no longer precisely circular as it is in afreshly formed fiber. Rather, the softening of the fibers and fusing ofadjacent fibers to each other imparts an irregular and irregularlyrounded shape to the fibers in transverse cross section. The transversecross sections here reflect joined fiber cross sections ranging inwidth—loosely, diameter—from about 50 to about 300 microns.

The fibers in the scaffold are bonded together and therefore are notloose fibers; but they retain their identities as separate fibers asshown in FIG. 2, which is a photograph of a longitudinal cross sectionof a scaffold (heated at 700° C.) of the invention. In one aspect of theinvention, at least about 75 or 85% by volume of the fibers in thescaffold are longitudinally co-aligned. In this regard the fibers areco-aligned longitudinally, where “co-aligned longitudinally” and thelike phrases (e.g., “in lengthwise co-alignment”) as applied to a groupof adjacent, bundled, or joined fibers in this application means thatthe alignment of each fiber in the group at any one place along at leastabout 75% of its length does not deviate more than about 25 degrees fromparallel to the central axis of the scaffold. In one preferredembodiment, each fiber in the group at any one place along at leastabout 75% of its length does not deviate more than about 15 degrees fromparallel to the central axis of the scaffold. In another preferredembodiment, each fiber in the group at any one place along at leastabout 75% of its length does not deviate more than about 10 degrees fromthe central axis of the scaffold. So it is evident that thisco-alignment aspect does not require 100% precise co-alignment of allfibers. The longitudinal co-alignment aspect also allows for some minordeviation of specific segments of individual fibers to an orientationoutside these 25, 15, and 10 degree requirements. This is reflected inthe requirement that the longitudinal co-alignment is of each fiberalong at least 75% of its length, rather than necessarily along itsentire length. So up to about 25% of the length of an individual fibermay be misaligned because, for example, it was bent during thescaffold-making process or otherwise. It can be seen therefore in FIG. 2that each fiber in the scaffold is not absolutely straight, nor is itlying along an absolutely straight line strictly parallel to all otherfibers in the scaffold. And each fiber is oriented generally in the samedirection, but each is not oriented in exactly the same direction.Moreover, the scaffold itself in certain embodiments is curved, bent, orotherwise not straight, in which cases the central axis of the scaffoldto which the alignment of the fibers is within 25 degrees of parallel isalso curved, bent, or otherwise not straight. It will also be evidentthat in certain embodiments a straight or curved scaffold will bemachined into a more complex shape as in FIGS. 7A and 7B, in whichinstance the scaffold central axis refers to the central axis as moldedand prior to machining.

In order to allow capillary action and channel-forming, the scaffoldtheoretically contains at least three fibers, although from FIG. 2 itcan be seen that the scaffold typically comprises dozens and evenhundreds of fibers. It can also be seen that the fibers lie generallylengthwise of the scaffold central axis A (i.e., lie generally in thedirection of the central axis) and are generally free of helicalorientation about the scaffold central axis. This arrangement applies toat least about 75 vol % of the fibers and preferably to substantiallyall of the fibers. The fibers shown here extend generally parallel tothe scaffold central axis A, which is also illustrated as axis B in FIG.3C, and axis C in FIG. 7B. This embodiment also manifests an optionalfeature that at least about 75 vol % of the fibers occupies the entirelength of the scaffold; but in other embodiments this is not the case.

The requirement of the invention that the fibers are co-alignedlongitudinally contemplates that the fibers are positioned so that theyhave a similar alignment, which similar alignment may be straight, bent,or curved. In a separate and distinct aspect of certain preferredembodiments, this common alignment is limited to a generally straightalignment along at least about 75%, 85%, or 95% of the length of thefibers. In other words, at least about 75%, 85%, or 95% of each fiber isgenerally straight, i.e., at least about 75%, 85%, or 95% of the lengthof each fiber has an alignment which is within 10 degrees of a meanstraight central axis for the fiber. So up to 5%, 15%, or 25% of thelength of each fiber may be curved, bent, or otherwise deviate more than10 degrees from straight in relation to the overall fiber length, butthe rest of each fiber is generally straight in that it so deviates lessthan 10 degrees. In one preferred embodiment, substantially the entirelength of each fiber is generally straight in that it deviates less than10 degrees from the fiber's average central axis. The “mean straightcentral axis” is the imaginary central axis for the fiber which isabsolutely straight and is an average of all axes along the fiberlength.

The fibers in the scaffold are selected to have characteristics suitablefor the specific application. In one embodiment, the fibers have alength between about 6 mm and about 15 cm, such as between about 12 mmand about 10 cm or between about 25 mm and about 75 mm. Each fiber has alength which is at least about 10 times its diameter. “Diameter” as usedherein refers to the fibers largest dimension transverse to its length,and it does not imply that the fibers are perfectly circular in crosssection. Each fiber therefore has a fiber lengthwise dimension which isat least about 10 times the fiber transverse dimension, e.g., diameter.In one embodiment, the fiber length is selected so that all,substantially all, or at least about 85 vol % of the individual fibersextend the entire length of the scaffold. The fibers may be selected tohave a pre-molding, pre-joining length which corresponds to the lengthof the scaffold. Or in most embodiments, the length of the fibers islonger than the desired ultimate scaffold length, and the scaffold iscut to the desired length after molding and joining. In an alternativeembodiment, the length of a substantial portion (e.g., at least 40 vol%) or all of the fibers is significantly less than the entire length ofthe scaffold.

FIGS. 1A, 1B, 1C, 4A and 4B also demonstrate the open and interconnectedporosity of the scaffold of the invention. The scaffold is manufacturedto have a sufficiently high open and interconnected porosity from end toend of the scaffold to facilitate capillary flow of fluids such asbodily fluids and medicines and components they carry through the lengthof the scaffold, as well as generally transverse from outside walls ofthe scaffold into the scaffold interior in directions generallytransverse to the longitudinal dimension of the fibers. And the scaffoldis manufactured so that the ultimate porosity is low enough that thescaffold has required strength for handling, implantation, and serviceafter implantation. If the porosity is too high, the scaffold risksbreakage in service, depending on where it is implanted and the loads itencounters. In a preferred embodiment, the porosity as measured involume is between about 10% and about 35%, for example between about 10%and about 30%, or between about 10% and about 25%. The porosity iscontrollable mainly by controlling the degree of softening of thefibers, in that highly softened fibers fuse together more completely toa structure with lower porosity. The degree of softening and fusing iscontrolled by controlling the joining temperature and time. Porosity isalso affected by the fiber diameter and by the range in fiber diameterwithin a given scaffold. Porosity tends to increase with an increasingrange in fiber diameter.

The scaffold of the invention in certain preferred embodiments for usein bone repair has a compressive strength between about 20 and about 250MPa, for example between about 20 and about 180 MPa or between about 80and about 140 MPa.

The fibers used in many embodiments of the invention are glass whereglass is defined as being at least 99 wt % an amorphous ornon-crystalline solid, for example made by fusing a mixture of oxidessuch as SiO₂, B₂O₃, P₂O₅ (known as glass forming oxides) with basicoxides such as the alkali and alkaline earth oxides. In an alternativeembodiment, the fibers include glass ceramics fibers that contain bothglassy and crystalline regions which in many respects function in thesame manner as a fiber that is completely (100%) non-crystalline. It isacceptable in some applications if the glass fiber crystallizes duringthe bonding step. The fibers may alternatively be pre-reacted bioactiveglasses such as glass fibers pre-reacted to have a thin surface layer ofhydroxyapatite. These foregoing different types of fibers are within acommon group which are referred to herein as “glass fibers.” In afurther alternative, the unidirectional scaffold comprises crystallinefibers (such as titanium wires) that would also provide a high amount ofcapillary action. Alternatively, the scaffold comprises a mix ofdifferent types of fibers selected from among these.

The fibers are preferably made from a material which is inorganic andwhich is biocompatible in that the fibers do not have adverse effectswhen implanted into mammals. Biocompatible materials include bothbioactive and bioinert materials. In certain preferred embodiments, thefibers are also of a bioactive glass in that they are of a glassmaterial which reacts with phosphorus such as phosphorus in bodilyfluids to form hydroxyapatite. Bioactive glasses are known in the art,for example, from U.S. Pat. No. 6,054,400; the entire disclosure ofwhich is incorporated herein by reference. Bioactive glasses areavailable, for example, from Mo-Sci Corporation of Rolla, Mo. In otherembodiments, some or all of the fibers may be bioinert rather thanbioactive, such as 100% bioinert fibers or a roughly 50/50 mix ofbioinert and bioactive fibers.

In general, bioactive glass is one which contains calcium and, whenplaced in contact with natural body fluids or simulated body fluids,forms a biocompatible calcium phosphate compound such as hydroxyapatite.When such a glass is immersed in or otherwise contacted with natural orsimulated body fluids which contain phosphate ions such as in a mammal,the glass dissolves, thereby releasing Ca²⁺ ions into the solution. Inthis solution, Ca²⁺ ions react with PO₄ ³⁻ and OH⁻ ions to form acalcium phosphate which has a relatively low solubility limit in theaqueous phosphate solution. As the dissolution of the glass proceeds,the concentration of calcium phosphate increases in the solution untilthe solubility limit of calcium phosphate is exceeded and, as aconsequence, hydroxyapatite (a form of calcium phosphate) is depositedas a porous layer on the outer surface of the dissolving glass. Theformation of this porous hydroxyapatite layer on the glass surface,however, does not prevent further dissolution of the glass. Rather, theglass continues to dissolve and, as it does, the thickness of the poroushydroxyapatite layer increases. Eventually, the glass is completelyreacted or transformed, leaving only a porous hydroxyapatite substancewhose shape and size are the same as the initial glass fiber.Hydroxyapatite has crystallographic and chemical properties similar tothose of mammalian bone. For example, human bone is a composite offibrous protein, collagen, and hydroxyapatite.

The material for use in the invention is also selected to be of acomposition which is available in fibers or which can be pulled intofibers. Glass fibers can be made several ways. For example, glass fiberscan be made by pulling by hand or with use of a rotating drum directlyfrom a melt, or they can be pulled through a bushing of a particularsize. The composition is preferably selected to be of a type whichsoftens to facilitate self-joining at a temperature below itscrystallization temperature. Suitable bioactive glasses include, forexample those listed in Table 1.

TABLE 1 Weight Percent Composition of Bioactive Glasses Li₂O Na₂O K₂OMgO CaO B₂O₃ P₂O₅ SiO₂ 0 20 10 5 10 0 0 55 0 18 9 0 14 1 4 54 0 12 15 511 1 2 54 0 6 12 5 20 0 4 53 0 18 6 2 17 2 2 53 0 15 12 2 11 3 4 53 0 2010 2 10 3 3 52 0 20 10 5 10 3 0 52 0 25 5 2 10 3 3 52 0 15 15 2 15 3 050 0 6 12 5 20 17.7 4 35.3 0 6 12 5 20 35.3 4 17.7 0 6 12 5 20 53 4 0 021.5 0 0 21.5 0 4 53 11.5 0 0 0 10 78.5 0 0 10.7 0 0 0 15 74.3 0 0 10 00 0 20 70 0 0

Glasses which crystallize under fiber-pulling conditions and/or whichcrystallize at a temperature below that where they soften sufficientlyfor joining such as 45S5 have some limited applications here but aregenerally avoided in the preferred embodiments because they become toobrittle and weak. Bioactive glasses such as 45S5 and other glasses thatcrystallize quickly not allowing sufficient self-bonding to occur may bebonded with sodium silicate or some other bonding agent to form analternative scaffold embodiment of the invention; however the strengthwill likely be relatively low in comparison with self-bonded scaffolds.

In forming the scaffold of the invention, a bundle of glass fibers suchas the 6.25 cm long loose cut fibers shown in FIG. 3A is placed as shownin 3B in a mold or similar vessel which, upon softening, joining, andcooling of the glass, will impart the desired final shape and strengthto the scaffold. Each of the bundles inserted as shown in FIG. 3B weighsabout 2.4 grams and the mold is about 5.6 cm long. In one embodiment,this vessel is a graphite mold such as a hollowed out cylinder as shownin 3B. The fibers are placed in the vessel tightly enough to fill thevessel cavity, but not so tightly as to risk breakage of the fibers orexcessive densification. The vessel is then placed in a furnace andheated at a rate of about 20° C./min in the presence of a suitableatmosphere such as air, oxygen, or nitrogen. The temperature and theheating time are selected depending on the glass composition to achievesoftening and bonding of the fibers while avoiding too much bondingwhich would not achieve the desired porosity. The joining is preferablyself-joining in that the softening of the glass accomplishes joining andno added joining agents are employed. That is, the scaffold bodyconsists only of joined fiber elements and no other elements. As ageneral proposition, the vessel is heated to a temperature between about500 and about 800° C. and held at that temperature for between about 5and about 60 minutes. For example, in one embodiment where the scaffoldis formed from type 13-93 glass fibers to a finished dimension of about62.5 mm long and about 6 mm in diameter, the vessel is heated to atemperature between about 695 and about 705° C. and held at thattemperature for between about 5 and about 45 minutes. After bonding, thevessel and scaffold are cooled, preferably in air, at a rate whichavoids cracking of the bonded fibers, such as between about 10 and about30° C. per minute.

The scaffold is then removed from the vessel and cut to the desiredlength to yield the product shown in FIG. 3C. Cutting is accomplished,for example, by filling the pores with a wax, cutting the scaffold tolength with a sharp, non-burring (e.g., diamond) saw, and thenchemically or thermally removing the wax. Sharp corners and edges areavoided by making clean cuts while the scaffold is impregnated with wax,or a polish can be done with either grinding paper or a mechanicalpolishing device such as a Dremel tool, also done while wax impregnated.

The scaffold can be pre-reacted in a phosphate solution such assimulated body fluid (SBF) or an alkali phosphate solution to form areacted surface layer of hydroxyapatite, prior to sterilization andimplantation in a mammal. The hydroxyapatite surface layer thickness canbe controlled by predetermined conversion kinetics of the glass in aphosphate containing solution. Heat treatment of the glass can induceglass crystallization which may be beneficial in the formation ofglass-ceramics or ceramics. Chemical (acid) etching may add surfaceroughness which could be beneficial to cell attachment. Heat treatingthe glass to cause phase separation to form multiple phases which couldreact at different rates and form a new microstructure within theindividual self-bonded fibers is desirable in certain applications. Itis also within the scope of this invention to incorporate additives suchas growth factors, medicines, etc. into the scaffold body which performa function such as assist with tissue regrowth or supplementreinforcement of the body. In most preferred embodiments, such additivesor reinforcements constitute less than about 10 vol. % of the scaffold,such that the fibers and the porosity cumulatively constitute at leastabout 75 vol. % of the scaffold body, for example at least about 90 vol.%. And in some embodiments there are no such additives orreinforcements, such that the scaffold body consists essentially of thefibers and the porosity.

After the wax has been removed, the scaffolds are sterilized. Apreferred method among several possible is dry heat sterilization. Thescaffolds are placed in a clean glass vial, covered with a cleanaluminum foil cap, and heated to approximately 300° C. for three to fourhours. Upon cooling, the sterile scaffolds are ready to be implanted.

Growth factors, medicines such as antibiotics, seeded cells or otherbiological material, liquids or gels of any type, coatings (particles,spheres, hollow spheres, thin film(s), fibers, and hollow fibers), aninterpenetrating phase such as a biodegradable polymer or bone cement(PMMA) or other biological polymer, other organic or inorganicmaterials, or any combination may be added after sterilization topromote the growth of tissues into the scaffold. Additionalsterilization may be required for scaffolds that have had inorganicnon-sterile components added, and the method of sterilization may varywith the material(s) added.

In one alternative embodiment of the invention, a titanium or otherbiocompatible support such as a rod is incorporated into the scaffold toprovide additional mechanical strength, as shown in FIGS. 10-13.

The unidirectional scaffold of the invention is suitable in one aspectfor forming a complete replacement bone or tissue segment where themammal's original bone or other tissue segment has been removed,crushed, decimated by disease or the like. In another aspect thescaffold is suitable as a bridge such as between about 2 mm and about 25mm in length for bridging two separated bone segments. The scaffold isintended to serve as a temporary bridge for facilitating fluidic (e.g.,marrow) communication (or transport) between the separated bonesegments, thereby facilitating the healing of the broken bones. Thescaffold also serves as an internal splint providing support for thebone fracture while the bone heals.

Example 1

Fibers of the bioactive glass type 13-93 (˜2.4 grams) produced atMissouri University of Science & Technology having diameters in therange of about 50 to about 400 microns and lengths of about 62 mm wereplaced inside a graphite mold formed by hollowing out a graphitecylinder. The mold was then placed in a furnace (Neytech Model 2-525)and heated in air to a temperature of about 700° C., where it was heldfor times ranging from 5 to 45 minutes. The heat source was discontinuedand the mold cooled to room temperature at an average cooling rate ofabout 30° C./min. Cylindrical scaffolds of unidirectional self-bondedfibers were removed from the mold, sectioned, and photographed toprovide the images in FIGS. 1A, 1B, and 1C. The diameter of thescaffolds was 6 mm. FIG. 1A depicts a scaffold heated for a shorterperiod of time having a lower degree of self bonding, higher porosity,and lower strength. FIGS. 1B and 1C depict a scaffold heated for alonger period of time, having a greater degree of self bonding, lowerporosity, and greater strength. In 1B and 1C the pore size isapproaching too small, thereby inhibiting fluid flow in the scaffold incomparison to FIG. 1A. Open pores of at least about 100 microns in crosssection are required for bone growth.

Example 2

A 6 mm diameter by 20 mm thick section of a unidirectional scaffoldproduced according to Example 1 was placed in a culture of osteoblastcells for four hours. MTT labeling was then performed on the section andphotographs were taken (FIGS. 4A and 4B). The dark spots are due to theuptake of viable osteoblast-like cells into the scaffold.

Example 3

Undirectional glass scaffolds were prepared in accordance with Example 1with joining/heating times at 700° C. of 5, 15, 30, and 45 minutes. Theopen porosity of each scaffold was then determined by the Archimedesliquid displacement method to be 35, 25, 15, and 18 vol %, respectively;see FIG. 5. This degree of open porosity is within the range of humancortical bone and was achieved for scaffolds heated for 15, 30, and 45minutes.

Example 4

Unidirectional glass scaffolds were prepared in accordance with Example1 with joining/heating times at 700° C. of 5, 15, 30, and 45 minutes.The average compressive strength of each scaffold was then determined bymechanical compression testing (Instron mechanical test instrument Model4204 with a crosshead speed of 0.5 mm/min) to be 20, 80, 130, and 112MPa, respectively, see FIG. 6. This demonstrates that compressivestrengths within the range of human cortical bone were achieved for thescaffolds heated for 30 and 45 minutes.

Example 5

A unidirectional scaffold was prepared in accordance with this inventionby molding, bonding, cooling, and then machining to mimic theconfiguration of a chicken bone. FIG. 7 shows the scaffold next to legbone of a chicken. The scaffold of the invention, which is thelighter-colored of the two specimens closer to the ruler in FIG. 7, hasa length of about 50 mm. This unidirectional scaffold is composed offibers which were oriented parallel to the longitudinal axis of thisobject.

Example 6

A unidirectional cylindrical glass scaffold was prepared in accordancewith Example 1 having a length of about 62 mm and a diameter of about 6mm. The fibers were type 13-93 bioactive glass having a length of about62 mm and diameters ranging from about 50 to about 400 microns. Thefibers were oriented parallel to the longitudinal axis of the scaffold.The tip of the scaffold was dipped in a glycerol solution (34 wt %glycerol-66 wt % distilled water) as shown in FIG. 8. This solution hasa viscosity of 2.5 centipoises at 25° C., which is in the average rangefor human blood. The purpose of this experiment was to demonstrate thestrong capillary forces which this scaffold exerts upon a liquidresembling human blood.

A roughly 40 second video of the experiment was filmed, and frames at 9,11, 15, and 22 seconds are shown in FIGS. 8A, 8B, 8C, and 8D,respectively. These frames show progressively upward darkening of thescaffold, which demonstrates rapid capillary uptake of the solution andits components into the scaffold, and that the scaffold of the inventionhas strong capillary uptake of fluid in the direction of thelongitudinal axis of the fiber scaffold.

Example 7

A cylindrical, unidirectional glass scaffold was prepared in accordancewith Example 1 having a length of about 62 mm and a diameter of about 6mm. The fibers were type 13-93 bioactive glass fibers having a length ofabout 62 mm and a diameter of about 50 to 400 microns. The fibers wereoriented parallel to the longitudinal axis of the scaffold.

An eyedropper was used to drop the same water-glycerol solution asdescribed in Example 6 onto the external surface of the scaffold asshown in FIG. 9 for the purpose of demonstrating the rapid capillaryaction of the scaffold in a direction perpendicular to the longitudinalaxis of the scaffold. The solution was dropped fairly quicklydrop-by-drop onto the external surface of the scaffold until thescaffold became saturated with the solution. A video of this experimentwas made over a 50 second period. Frames at 4 seconds (0 drops), 12seconds (3 drops), 18 seconds (6 drops), and 43 seconds (19 drops) areshown in FIGS. 9A, 9B, 9C, and 9D, respectively. These frames show rapidcapillary uptake of the solution and its components into the scaffold inboth the lengthwise and transverse directions This experimentdemonstrates the scaffold's high affinity for the liquid and that thescaffold retained most of the drops before becoming saturated. At theconclusion of this experiment most of the liquid was retained in thescaffold and very little of the solution had dripped out of the scaffoldinto the bowl, as shown in 9D.

Example 8

A unidirectional scaffold was prepared generally in accordance withExample 1 having a length of about 62 mm and a diameter of about 6 mm.The fibers were type 13-93 bioactive glass fibers having a length ofabout 62 mm and a diameter of about 50 to 400 microns. The fibers wereoriented in longitudinal co-alignment defining the length of thescaffold with an added reinforcing rod placed at the center of thefibers prior to self-bonding. FIG. 10 is a schematic pictorialdemonstrating this concept of placing a reinforcement of Ti or othermetal or alloy or supporting material in the center of a self-bondedunidirectional bioactive glass fiber scaffold. FIG. 11 is a photographof the reinforced unidirectional bioactive glass scaffold prepared witha Ti rod placed in the center of a bundle of fibers, and heated so thatthe fibers self-bonded and also attached to the rod, in accordance withthis example.

FIG. 12 is a schematic pictorial demonstrating an alternative concept offilling a hollow reinforcing tube of Ti or other metal or alloy orpolymer including biodegradable polymers such as PCL or PLA or othersupporting material with unidirectional glass fiber followed byself-bonding to form the scaffold. The tube is, for example, made oftitanium or any other reinforcing material with similar thermalexpansion properties of the glass so the scaffold bonds to the tube asthe fibers self-bond. This may be beneficial in promoting bone ingrowthin prosthesis such as hip implants by inserting glass fibers into andaround the implant as a means for improved bone attachment and ingrowth.FIG. 12 demonstrates an aspect of the invention, as with FIGS. 1A, 1B,and 1C, in that the fibers extending lengthwise of the scaffold centralaxis define channels lengthwise through a core within the scaffold. Thisis distinct from the alternative in FIG. 11, in which the core isoccupied by a reinforcing rod rather than by channel-defining fibers.

FIG. 13 is a schematic pictorial demonstrating a further alternativeconcept of a unidirectional bioactive glass scaffold filled with apolymer phase. Suitable polymers include those such as bone cement(PMMA) or biodegradable polymers such as PCL or PLA, as the polymerphase is used for sustained reinforcement (PMMA) or an initialreinforcement followed by a slow degradation of biodegradable polymerallowing new tissue to fill in with time. These methods associated withFIGS. 10-13 can be practiced individually or in any combination forconstructing or implementing a reinforced unidirectional bioactive glassscaffold. These embodiments are distinct from the embodiments featuredin FIGS. 8 and 9, where the scaffold body consists essentially of thefibers and porosity as described herein.

Example 9

FIG. 14 is a schematic illustration of an alternative embodiment of theinvention in which biocompatible glass fibers longitudinally aligned inaccordance with the invention are bonded together by dipping into amolten polymer to form a rigid outer layer, depicted as black in thefigure. FIG. 15 is a photograph of a scaffold prepared this way bydipping a bundle of type 13-93 fibers into molten polymer of 82 partspolylactic acid (PLA) and 18 parts polyglycolic acid (PGA) at 250° C.The degradation rate can be controlled by selecting a mix ofbiocompatible polymers in proportions to impart the desired degradation.In this embodiment and other embodiments, it is also possible to sprayor otherwise coat the individual fibers with a polymer such as PLA orPGA which affects degradation.

Example 10

FIG. 16 is a schematic illustration of an alternative embodiment of theinvention in which biocompatible glass fibers longitudinally aligned inaccordance with the invention are bonded together by dipping into a bathof a polymer dissolved in a solvent. By dissolving the polymer in asolvent, the polymer depicted here in black is able to flow into theinterior of the bundle of fibers, in contrast to the embodiment in FIG.14. FIG. 17 is a photograph of a scaffold prepared this way by dipping abundle of type 13-93 fibers into a polymer of 82 parts polylactic acid(PLA) and 18 parts polyglycolic acid (PGA) dissolved in a solvent.

Example 11

FIG. 18 is a schematic illustration of a further alternative embodimentof the invention in which biocompatible glass fibers longitudinallyaligned in accordance with the invention are precoated with PLA, thenbonded together as shown in FIG. 19. FIG. 20 is a photograph of ascaffold prepared this way by dipping a bundle of type 13-93 fibers intoan 82:18 mixture of PLA/PGA in a solvent. In this embodiment, thethickness of the polymer coating is, for example, at least about 500 nm,such as between about 0.5 micron and about 10 microns, or between about1 micron and about 5 microns.

It can be seen from FIGS. 14-20, therefore, that the rigidity requiredin the invention can be imparted by bonding the longitudinally alignedfibers to each other by polymer bonding. As further alternatives, theprecoated longitudinally aligned fibers shown in FIG. 18 can be bondedto each other by exposing the polymer coating to a solvent to partiallydissolve the coating. Then adjacent fibers bond to each other as thepolymer re-solidifies upon removal of the solvent. Or the polymercoating on the fibers can be softened by heating, then hardened bycooling, leaving adjacent fibers bonded to each other. A still furtheralternative is to dust the fibers with polymer such that the polymer ismore occasional and intermittent as shown in FIG. 21, in contrast to thecontinuous polymer coating shown in FIG. 18. Then the scaffold can beheated to soften the polymer such that it will bond the adjacentlongitudinally aligned fibers at various points.

Among the polymer materials suitable for the polymer tube shown in FIG.12, the polymer fill in FIG. 13, the polymer dip coating in FIG. 14, andthe polymer coating on fibers in FIG. 18 are, for example,polycaprolactones (PCL), poly-L-Lactic acid (PL-LA), polyvinyl alcohol(PVA), polyglycolic acid (PGA), polyacrylic acids (PAA), poly ethyleneglycol (PEG), poly-L-Lactic gylcolic acid (PLGA), polyesters,polyalkenoics, polyolefins, polysulfones, poly(anhydrides), poly(hydroyacids), polyglycolides, polylactides, poly(propylene fumerates),polyacetals, polycarbonates, polyamino acids, poly)orthoesters),polyamides, poly(vinyl pryyolidones), poly(dioxanones),polyhydroxyvalyrates, polyhydroxybutyrates, biodegradablepolycyanoacrylates, biodegradable polyurethanes, poly(methyl vinylether), poly(esteramides), polyketals, poly(glyconates), poly(maleicanhydride), poly(maleic acid), poly(alkylenesuccinates), poly(ppyrrole),polyphosphazines, poly(maleic anhydride), tyrosine-based polymers,polysaccharides, poly(alkylene oxalates), poly(orthocarbonates),poly(ethylene oxide), polyureas, poly(ethylene vinyl acetate),polystyrene, polypropylene, polymethacrylate, polyethylene,poly(aniline), poly(thiophene), non-biodegradable polyurethanes,co-polymers, adducts, and mixtures thereof.

In view of the above, it will be seen that the several objects of theinvention are achieved and other advantageous results attained.

When introducing elements of the present invention or the preferredembodiments(s) thereof, the articles “a”, “an”, “the” and “said” areintended to mean that there are one or more of the elements. The terms“comprising”, “including” and “having” are intended to be inclusive andmean that there may be additional elements other than the listedelements.

As various changes could be made in the above compositions and methodswithout departing from the scope of the invention, it is intended thatall matter contained in the above description and shown in theaccompanying drawings shall be interpreted as illustrative and not in alimiting sense.

The invention claimed is:
 1. A tissue scaffold for repair andregeneration of bone hard tissue or muscle, skin, or organ soft tissue,the scaffold comprising: a rigid scaffold body having a scaffold centralaxis, a scaffold transverse dimension, and a scaffold lengthwisedimension which is greater than the scaffold transverse dimension, thescaffold body having a compressive strength between about 20 and about250 MPa and comprising: biocompatible inorganic glass fibers each havinga fiber transverse dimension and a fiber lengthwise dimension which isat least about 10 times the fiber transverse dimension; and wherein eachof the fibers has a diameter between about 20 and about 5000 microns;wherein at least about 75 vol % of the fibers are longitudinallyco-aligned and lie generally lengthwise of the scaffold central axis,are generally free of helical orientation about the scaffold centralaxis, and are arranged to define open channels within the scaffold whichallow fluid flow into and lengthwise within the scaffold; and whereinadjacent longitudinally co-aligned fibers are bonded together.
 2. Thetissue scaffold of claim 1 wherein each fiber among the at least about75 vol % of fibers has an alignment along at least about 75% of itslength which does not deviate more than about 25 degrees from parallelto the central axis of the scaffold.
 3. The tissue scaffold of claim 1wherein each fiber among the at least about 75 vol % of fibers isgenerally straight.
 4. The tissue scaffold of claim 1 wherein thescaffold body has an interconnected porosity.
 5. The tissue scaffold ofclaim 4 wherein the fibers and the porosity cumulatively constitute atleast about 75 vol. % of the scaffold body.
 6. The tissue scaffold ofclaim 4 wherein the scaffold body consists essentially of the glassfibers and the porosity.
 7. The tissue scaffold of claim 1 wherein thescaffold body has an interconnected porosity between about 10 vol. % andabout 35 vol. % of the scaffold body.
 8. The tissue scaffold of claim 5wherein the scaffold body consists essentially of the glass fibers andthe porosity.
 9. The tissue scaffold of claim 1 wherein the fibers arebioactive glass fibers.
 10. The tissue scaffold of claim 1 wherein thefibers are self-bonded together in that adjacent longitudinally alignedfibers are fused together.
 11. The tissue scaffold of claim 1 whereinthe scaffold body of fibers is filled with a polymer phase.
 12. Thetissue scaffold of claim 11 wherein the polymer phase comprises PMMA,PCL, or PLA.
 13. The tissue scaffold of claim 11 wherein the polymerphase is a polymer selected from the group consisting ofpolycaprolactones (PCL), poly-L-Lactic acid (PL-LA), polyvinyl alcohol(PVA), polyglycolic acid (PGA), polyacrylic acids (PAA), poly ethyleneglycol (PEG), poly-L-Lactic gylcolic acid (PLGA), polyesters,polyalkenoics, polyolefins, polysulfones, poly(anhydrides), poly(hydroyacids), polyglycolides, polylactides, poly(propylene fumerates),polyacetals, polycarbonates, polyamino acids, poly)orthoesters),polyamides, poly(vinyl pryyolidones), poly(dioxanones),polyhydroxyvalyrates, polyhydroxybutyrates, biodegradablepolycyanoacrylates, biodegradable polyurethanes, poly(methyl vinylether), poly(esteramides), polyketals, poly(glyconates), poly(maleicanhydride), poly (maleic acid), poly (alkylene succinates),poly(ppyrrole), polyphosphazines, poly(maleic anhydride), tyrosine-basedpolymers, polysaccharides, poly(alkylene oxalates),poly(orthocarbonates), poly(ethylene oxide), polyureas, poly(ethylenevinyl acetate), polystyrene, polypropylene, polymethacrylate,polyethylene, poly(aniline), poly(thiophene), non-biodegradablepolyurethanes, co-polymers, adducts, and mixtures thereof.
 14. Thetissue scaffold of claim 1 wherein the rigid scaffold body furthercomprises a reinforcing tube of biodegradable polymer with thebiocompatible inorganic glass fibers within the tube.
 15. The tissuescaffold of claim 14 wherein the biodegradable polymer comprises PCL orPLA.
 16. The tissue scaffold of claim 15 wherein the biodegradablepolymer is selected from the group consisting of polycaprolactones(PCL), poly-L-Lactic acid (PL-LA), polyvinyl alcohol (PVA), polyglycolicacid (PGA), polyacrylic acids (PAA), poly ethylene glycol (PEG),poly-L-Lactic gylcolic acid (PLGA), polyesters, polyalkenoics,polyolefins, polysulfones, poly(anhydrides), poly(hydroy acids),polyglycolides, polylactides, poly(propylene fumerates), polyacetals,polycarbonates, polyamino acids, poly)orthoesters), polyamides,poly(vinyl pryyolidones), poly(dioxanones), polyhydroxyvalyrates,polyhydroxybutyrates, biodegradable polycyanoacrylates, biodegradablepolyurethanes, poly(methyl vinyl ether), poly(esteramides), polyketals,poly(glyconates), poly(maleic anhydride), poly (maleic acid), poly(alkylene succinates), poly(ppyrrole), polyphosphazines, poly(maleicanhydride), tyrosine-based polymers, polysaccharides, poly(alkyleneoxalates), poly(orthocarbonates), poly(ethylene oxide), polyureas,poly(ethylene vinyl acetate), polystyrene, polypropylene,polymethacrylate, polyethylene, poly(aniline), poly(thiophene),non-biodegradable polyurethanes, co-polymers, adducts, and mixturesthereof.
 17. The scaffold of claim 1 wherein adjacent longitudinallyco-aligned fibers are bonded together by adhesive or by fusing at atemperature at which the fibers soften and self-bond to each other. 18.The tissue scaffold of claim 17 wherein the adjacent longitudinallyco-aligned fibers are bonded together by the adhesive, the adhesive is apolymer, and the adjacent longitudinally co-aligned fibers are bondedtogether by polymer bonding.
 19. The tissue scaffold of claim 18 whereinthe adjacent longitudinally co-aligned fibers have a coating of saidpolymer thereon.
 20. The tissue scaffold of claim 19 wherein the polymercoating comprises polymer selected from the group consisting of PLA,PGA, and mixtures thereof.
 21. The tissue scaffold of claim 19 whereinthe polymer coating comprises polymer selected from the group consistingof polycaprolactones (PCL), poly-L-Lactic acid (PL-LA), polyvinylalcohol (PVA), polyglycolic acid (PGA), polyacrylic acids (PAA), polyethylene glycol (PEG), poly-L-Lactic gylcolic acid (PLGA), polyesters,polyalkenoics, polyolefins, polysulfones, poly(anhydrides), poly(hydroyacids), polyglycolides, polylactides, poly(propylene fumerates),polyacetals, polycarbonates, polyamino acids, poly)orthoesters),polyamides, poly(vinyl pryyolidones), poly(dioxanones),polyhydroxyvalyrates, polyhydroxybutyrates, biodegradablepolycyanoacrylates, biodegradable polyurethanes, poly(methyl vinylether), poly(esteramides), polyketals, poly(glyconates), poly(maleicanhydride), poly (maleic acid), poly (alkylene succinates),poly(ppyrrole), polyphosphazines, poly(maleic anhydride), tyrosine-basedpolymers, polysaccharides, poly(alkylene oxalates),poly(orthocarbonates), poly(ethylene oxide), polyureas, poly(ethylenevinyl acetate), polystyrene, polypropylene, polymethacrylate,polyethylene, poly(aniline), poly(thiophene), non-biodegradablepolyurethanes, co-polymers, adducts, and mixtures thereof.
 22. Thescaffold of claim 19 wherein the bonding is achieved by dissolving andre-solidifying the polymer coating.
 23. The scaffold of claim 19 whereinthe bonding is achieved by softening by heating then hardening bycooling the polymer coating.
 24. The scaffold of claim 17 wherein theadjacent longitudinally co-aligned fibers are bonded together by theadhesive and the adhesive comprises an intermittent polymer coating onthe fibers.
 25. The scaffold of claim 1 wherein each of the fibers has adiameter between about 50 and about 400 microns.
 26. The scaffold ofclaim 1 wherein each of the fibers has a diameter between about 100 andabout 450 microns.